Increase in activity of glucose 6-phosphate dehydrogenase in mouse mammary tissue cultured in insulin.

Abstract

1. In organ cultures of mammary tissue from C3H mice we observed increases in the activity of glucose 6-phosphate dehydrogenase similar to that occurring at parturition. 2. In 22hr. cultures of tissue from late-pregnant mice insulin was required for the increases, but the further addition of prolactin, corticosterone and certain other hormones had no effect. The rise in activity occurred over the second half of the culture period. 3. Results from culture of adipose tissue, and mammary tissue rich in adipose tissue, strongly suggest that the rise in activity occurs in mammary parenchymal rather than adipose cells. 4. In 45hr. cultures prolactin prevented a fall in enzyme activity between 22hr. and 45hr. If the medium contained serum the activity at 22hr. was unaffected, but it continued to rise up to 45hr., and prolactin then had no effect. 5. The enzyme also increased in activity in cultures of mammary tissue from mid-pregnant mice. Insulin was again required, the activity was higher at 45hr. than at 24hr. and prolactin increased the activities at both these times. 6. Actinomycin D, cycloheximide and puromycin at low concentration in the media of 22hr. cultures all prevented increases in enzyme activity. Hydroxyurea at a concentration that inhibited the incorporation of [(3)H]thymidine into DNA by 92% had little effect. 7. Actinomycin D and cycloheximide largely failed to prevent the rise in enzyme activity if added after 3.5hr. and 12hr. respectively. Hence all essential RNA and protein synthesis appears to be finished by 3.5hr. and 12hr., although most of the increase in enzyme activity occurs gradually between 12hr. and 22hr. 8. We suggest that the increases in enzyme activity, both in culture and in the living animal at parturition, are induced by an influx of glucose that is restrained during pregnancy by the growth-hormone-like action of placental lactogen.